TABLE 6.
Primers used in this study
| Name | Commentsb | Sequence (5′–3′)a |
|---|---|---|
| TC1 | Cloning IRC7S or IRC7L using EcoRI (underlined), forward | GACTGAATTCATGATTGATCGTACCGAGTTATCG |
| TC2 | Cloning IRC7S using XhoI (underlined), reverse | TAGCCTCGAGCTAGCCACCCCATGAAAATCCCA |
| TC3 | Cloning IRC7L using XhoI (underlined), reverse | GACTCTCGAGTCAGATTTGTAAAGGATTCAAAG |
| TC4 | Sequencing primer (pHVX2 plasmid), forward | TCAAACAGAATTGTCCGAATCG |
| TC5 | Sequencing primer (pHVX2 plasmid), reverse | CCGAACATAGAAATATCGAATGG |
| TC6 | Site-directed mutagenesis (IRC7L K43R), forward | GGGGTCAACCATCATTCTTAgAAAACTTAGTGATTTAGAAC |
| TC7 | Site-directed mutagenesis (IRC7L M106I), forward | GCTGGTGATCATATCTTGATtACTGATAGTGTCTACGTGC |
| TC8 | Site-directed mutagenesis (IRC7L P146R), forward | GATATAGAAAAACTAGTTAAGCgAAATACAACCGTCATTTTCCTC |
| TC9 | Site-directed mutagenesis (IRC7L G253R), forward | GTCAATTAGCATTGCGAaGAATGCGTACATTGCAC |
| TC10 | Site-directed mutagenesis (IRC7L W274R), forward | CCTGGATTTGGCTGCTcGGCTCGGAAATCGAGAT |
| TC11 | Site-directed mutagenesis (IRC7L G321D), forward | GAGGAAAATGACGGTTGTATTTcGCTTAACTAGTTTTTCTATATC |
| TC12 | Site-directed mutagenesis (IRC7L E323G), forward | CACAAGAGCTGGTCTGGgGAAAATGGTAGAAGGGA |
| TC13 | Deletion of IRC7 using CORE cassette, forward | ATGATTGATCGTACCGAGTTATCGAAGTTTGGTATTACTACGCAACTGTCgagcccccgatttagagctt |
| TC14 | Deletion of IRC7 using CORE cassette, reverse | TCAGATTTGTAAAGGATTCAAAGAAATTTCTTCTGCGAGACGTTCAAAGCCctcgaggtcgacggtatcgat |
| TC15 | Confirmation deletion IRC7 (5′-UTR IRC7), forward | ATTGTCGCCTCTTGTTTTGAG |
| TC16 | Confirmation deletion IRC7 (5′), reverse | AAGACTCTCCTCCGTGCGTC |
| TC17 | Confirmation deletion IRC7 (3′-UTR IRC7), reverse | GGAGTGAGATCACATGTCATATTG |
| TC18 | Confirmation deletion IRC7 (3′), forward | TGAGCATGCCCTGCCCCTA |
a
For primers TC6 through TC12, the reverse primers used for site-directed mutagenesis are the reverse complement of the corresponding forward primers; the nucleotide to be changed is indicated in lowercase in the sequence. For primers TC13 through TC14, the sequence annealing the CORE cassette of the pCORE5 plasmid is indicated in lowercase.
b
UTR, untranscribed region.